1. Field
The present application relates to an observing apparatus performing a time lapse shooting of an observation object such as a living sample and an observation method.
2. Description of the Related Art
Conventionally, a living sample observing apparatus performing a time lapse shooting while incubating cells in an incubation container is known in Japanese Unexamined Patent Application Publication No. 2004-309719.
On the other hand, in recent years, an art manufacturing, differentiating and inducing iPS cells (Induced Pluripotent Stem cell) from cultured cells differentiated into somatic cells is focused and various studies have been done in various fields such as a regenerative medicine field and a development of new drugs field.
As a representative method manufacturing the iPS cells from the cultured cells, a method in which an exogenous gene called as Yamanaka factors (Sox 2, K1f4, Oct3/4, c-Myc) is introduced into the cultured cells with a vector such as retrovirus and plasmid, and they are incubated for several weeks is known. The cells to which the exogenous gene is introduced are reprogrammed (initialized) and come to have equivalent characteristics as ES cells (a characteristic multiplying in a colony state mode, and having pluripotency) in various points. In the cells initialized as stated above, an endogenous gene peculiar to the iPS cells such as a Nanog gene is expressed. Besides, the expression of the introduced exogenous gene is terminated simultaneously with the initialization of cells caused by a mechanism called as a silencing in the cell.
In this method, it becomes important to monitor a process from the expression of the exogenous gene to the termination of the expression thereof caused by the silencing (1) and a process in which the endogenous gene peculiar to the iPS cells is expressed by the initialization (2) from points of view of an evaluation, an explanation of function of the iPS cells.
It is possible to visualize these processes (1) and (2), and for example, when the process (1) is visualized, a fluorescent protein gene (DsRed gene) which is expressed together with the expression of the above-stated exogenous gene (Sox2, K1 f4, Oct 3/4, c-Myc) and silencing thereof occurs together with the silencing of the exogenous gene is introduced into the cultured cells together with the exogenous gene. Besides, when the process (2) is visualized, the endogenous gene (Nanog gene) in experimental animals is recomposed into a gene accompanied by the fluorescent protein (Nanog-GFP gene) in advance.
However, manufacturing efficiency of the iPS cells is currently low, and therefore, the iPS cells are not expressed only at a very small part in the incubation container. Besides, it is currently impossible to predict a part where the iPS cells are expressed. Accordingly, it is difficult to effectively observe the processes (1) and (2) in which the iPS cells are initialized with the above-stated living sample observing apparatus.
A proposition of the present application is to provide an observing apparatus and an observation method capable of effectively observe a certain material which is irregularly expressed in an observation area.